Characterization of novel bacteriophage phiC119 capable of lysing multidrug-resistant Shiga toxin-producingEscherichia coliO157:H7

BackgroundShiga toxin-producingEscherichia coli(STEC) is one of the most common and widely distributed foodborne pathogens that has been frequently implicated in gastrointestinal and urinary tract infections. Moreover, high rates of multiple antibiotic-resistantE. colistrains have been reported worldwide. Due to the emergence of antibiotic-resistant strains, bacteriophages are considered an attractive alternative to biocontrol pathogenic bacteria. Characterization is a preliminary step towards designing a phage for biocontrol.MethodsIn this study, we describe the characterization of a bacteriophage designated phiC119, which can infect and lyse several multidrug-resistant STEC strains and someSalmonellastrains. The phage genome was screened to detect thestx-genes using PCR, morphological analysis, host range was determined, and genome sequencing were carried out, as well as an analysis of the cohesive ends and identification of the type of genetic material through enzymatic digestion of the genome.ResultsAnalysis of the bacteriophage particles by transmission electron microscopy showed that it had an icosahedral head and a long tail, characteristic of the familySiphoviridae. The phage exhibits broad host range against multidrug-resistant and highly virulentE. coliisolates. One-step growth experiments revealed that the phiC119 phage presented a large burst size (210 PFU/cell) and a latent period of 20 min. Based on genomic analysis, the phage contains a linear double-stranded DNA genome with a size of 47,319 bp. The phage encodes 75 putative proteins, but lysogeny and virulence genes were not found in the phiC119 genome.ConclusionThese results suggest that phage phiC119 may be a good biological control agent. However, further studies are required to ensure its control of STEC and to confirm the safety of phage use.

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Antimicrobial activity of Syzygium aromaticum L. essential oil on extended-spectrum beta-lactamases-producing Escherichia coli

Bulletin of the National Research Centre ◽

10.1186/s42269-020-00458-x ◽

2020 ◽

Vol 44 (1) ◽

Author(s):

E. L. Mejía-Argueta ◽

J. G. Santillán-Benítez ◽

M. M. Canales-Martinez ◽

A. Mendoza-Medellín

Keyword(s):

Escherichia Coli ◽

Essential Oil ◽

Genetic Material ◽

Gastrointestinal Diseases ◽

Syzygium Aromaticum ◽

Antibiotic Resistant ◽

Plant Essential Oils ◽

E Coli ◽

Extended Spectrum ◽

Tract Infections

Abstract Background To test the antimicrobial potential of clove essential oil that has been less investigated on antimicrobial-resistant organisms (extended-spectrum β-lactamase-ESBL-producing Escherichia coli), we collected 135 ESBL-producing Escherichia coli strains given that E. coli is the major organism increasingly isolated as a cause of complicated urinary and gastrointestinal tract infections, which remains an important cause of therapy failure with antibiotics for the medical sector. Then, in this study, we evaluated the relationship between the antibacterial potential activity of Syzygium aromaticum essential oil (EOSA) and the expression of antibiotic-resistant genes (SHV-2, TEM-20) in plasmidic DNA on ESBL-producing E. coli using RT-PCR technique. Results EOSA was obtained by hydrodistillation. Using Kirby-Baüer method, we found that EOSA presented a smaller media (mean = 15.59 mm) in comparison with chloramphenicol (mean = 17.73 mm). Thus, there were significant differences (p < 0.0001). Furthermore, EOSA had an antibacterial activity, particularly on ECB132 (MIC: 10.0 mg/mL and MBC: 80.0 mg/mL), and a bacteriostatic effect by bactericidal kinetic. We found that the expression of antibiotic-resistant gene blaTEM-20 was 23.52% (4/17 strains) and no expression of blaSHV-2. EOSA presented such as majority compounds (eugenol, caryophyllene) using the GC–MS technique. Conclusions Plant essential oils and their active ingredients have potentially high bioactivity against a different target (membranes, cytoplasm, genetic material). In this research, EOSA might become an important adjuvant against urinary and gastrointestinal diseases caused by ESBL-producing E. coli.

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JD419, a Staphylococcus aureus Phage With a Unique Morphology and Broad Host Range

Frontiers in Microbiology ◽

10.3389/fmicb.2021.602902 ◽

2021 ◽

Vol 12 ◽

Author(s):

Tingting Feng ◽

Sebastian Leptihn ◽

Ke Dong ◽

Belinda Loh ◽

Yan Zhang ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Host Range ◽

Antibiotic Resistance Genes ◽

Multidrug Resistant ◽

Complete Characterization ◽

Resistant Bacteria ◽

Broad Host Range ◽

Gene Engineering ◽

Mrsa Strains

Phage therapy represents a possible treatment option to cure infections caused by multidrug-resistant bacteria, including methicillin and vancomycin-resistant Staphylococcus aureus, to which most antibiotics have become ineffective. In the present study, we report the isolation and complete characterization of a novel phage named JD219 exhibiting a broad host range able to infect 61 of 138 clinical strains of S. aureus tested, which included MRSA strains as well. The phage JD419 exhibits a unique morphology with an elongated capsid and a flexible tail. To evaluate the potential of JD419 to be used as a therapeutic phage, we tested the ability of the phage particles to remain infectious after treatment exceeding physiological pH or temperature. The activity was retained at pH values of 6.0–8.0 and below 50°C. As phages can contain virulence genes, JD419’s complete genome was sequenced. The 45509 bp genome is predicted to contain 65 ORFs, none of which show homology to any known virulence or antibiotic resistance genes. Genome analysis indicates that JD419 is a temperate phage, despite observing rapid replication and lysis of host strains. Following the recent advances in synthetic biology, JD419 can be modified by gene engineering to remove prophage-related genes, preventing potential lysogeny, in order to be deployed as a therapeutic phage.

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Characterization of auto-agglutinating and non-typeable uropathogenic Escherichia coli strains

The Journal of Infection in Developing Countries ◽

10.3855/jidc.11098 ◽

2019 ◽

Vol 13 (06) ◽

pp. 465-472

Author(s):

Ulises Hernández-Chiñas ◽

Alejandro Pérez-Ramos ◽

Laura Belmont-Monroy ◽

María E Chávez-Berrocal ◽

Edgar González-Villalobos ◽

...

Keyword(s):

Escherichia Coli ◽

Virulence Genes ◽

Antimicrobial Agents ◽

Multidrug Resistant ◽

Uropathogenic Escherichia Coli ◽

E Coli ◽

Resistant Strains ◽

Tract Infections ◽

Disk Method

Introduction: Uropathogenic Escherichia coli (UPEC) are the main etiological agent of urinary tract infections (UTIs). Association between different serotypes and UTIs is known, however, some strains are incapable to be serotyped. The aim of this work was to study bthe phenotypical and genotypical characteristics of 113 non-typeable (NT) and auto-agglutinating (AA) E. coli strains, isolated from UTIs in children and adults. Methodology: The 113 UPEC strains were analyzed by PCR assays using specific primers to determine their serogroups, fimH, papC, iutA, sat, hlyCA and cnf1, virulence associated genes, and chuA, yjaA and TSPE4.C2 for phylogroup determination. Additionally, the diffusion disk method was performed to evaluate the antimicrobial resistance to 18 antimicrobial agents. Results: Using the PCR assay, 63% (71) of the strains were genotyped showing O25 and O75 as the most common serogroups. The virulence genes fimH (86%) and iutA (74%) were the most prevalent, in relation to the phylogroups the commensal (A and B1) and virulent (B2 and D) showed similar frequencies (P > 0.05). The antimicrobial susceptibility test showed a high percentage (73%) of multidrug-resistant strains. Conclusions: The genotyping allowed identifying the serogroup in many of the strains that could not be typed by traditional serology. The strains carried virulence genes and were multidrug-resistant in both, commensal and virulent phylogroups. Our findings revealed that, in addition to the classical UPEC serogroups, there are pathogenic serogroups not reported yet.

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Genomic Insights into a Colistin-Resistant Uropathogenic Escherichia coli Strain of O23:H4-ST641 Lineage Harboring mcr-1.1 on a Conjugative IncHI2 Plasmid from Egypt

Microorganisms ◽

10.3390/microorganisms9040799 ◽

2021 ◽

Vol 9 (4) ◽

pp. 799

Author(s):

Azza S. Zakaria ◽

Eva A. Edward ◽

Nelly M. Mohamed

Keyword(s):

Escherichia Coli ◽

Multidrug Resistant ◽

Coli Strain ◽

Colistin Resistance ◽

E Coli ◽

Agriculture Sector ◽

Potential Reservoir ◽

Clinical Source ◽

Tract Infections

The reintroduction of colistin, a last-resort antibiotic for multidrug-resistant pathogens, resulted in the global spread of plasmid-mediated mobile colistin resistance (mcr) genes. Our study investigated the occurrence of colistin resistance among Escherichia coli isolated from patients with urinary tract infections admitted to a teaching hospital in Egypt. Out of 67 isolates, three isolates were colistin-resistant, having a minimum inhibitory concentration of 4 µg/mL and possessing the mcr-1 gene. A double mechanism of colistin resistance was detected; production of mcr-1 along with amino acid substitution in PmrB (E123D and Y358N) and PmrA (G144S). Broth mating experiments inferred that mcr-1 was positioned on conjugative plasmids. Whole-genome sequencing of EC13049 indicated that the isolate belonged to O23:H4-ST641 lineage and to phylogroup D. The mcr-1-bearing plasmid corresponded to IncHI2 type with a notable similarity to other E. coli plasmids previously recovered from Egypt. The unbanned use of colistin in the Egyptian agriculture sector might have created a potential reservoir for the mcr-1 gene in food-producing animals that spread to humans. More proactive regulations must be implemented to prevent further dissemination of this resistance. This is the first characterization of mcr-1-carrying IncHI2:ST4 plasmid recovered from E. coli of a clinical source in Egypt.

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Complete Genome Sequence of Broad-Host-Range Shiga Toxin-Converting Bacteriophage SH2026Stx1, Isolated from Escherichia coli O157:H7

Genome Announcements ◽

10.1128/genomea.00490-18 ◽

2018 ◽

Vol 6 (25) ◽

Author(s):

Mingrui Duan ◽

Samuel S. Hunter ◽

Scott A. Minnich ◽

Matthew W. Fagnan ◽

Daniel D. New ◽

...

Keyword(s):

Escherichia Coli ◽

Host Range ◽

Genome Sequence ◽

Shiga Toxin ◽

Complete Genome Sequence ◽

Complete Genome ◽

Broad Host Range ◽

Escherichia Coli O157 ◽

Content Type ◽

E Coli

ABSTRACT The Shiga toxin-encoding phage SH2026Stx1 was isolated from Escherichia coli O157:H7 strain 2026. SH2026Stx1 and its detoxified derivative can infect a broad range of E. coli strains, including commensal, enteropathogenic, and enteroaggregative strains. We report here the complete genome sequence of phage SH2026Stx1 and its important features.

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Characterization of Tn7-like transposons and its related antibiotic resistance among Enterobacteriaceae from livestock and poultry in China

10.21203/rs.3.rs-20496/v1 ◽

2020 ◽

Cited By ~ 1

Author(s):

Juan He ◽

Cui Li ◽

Pengfei Cui ◽

Hongning Wang

Keyword(s):

Antibiotic Resistance ◽

Resistance Genes ◽

Horizontal Transmission ◽

Antibiotic Resistance Genes ◽

Variable Region ◽

Multidrug Resistant ◽

Antibiotic Resistant ◽

E Coli ◽

New Type

Abstract Background: This study was aimed to investigate the prevalence and structure of Tn7-like in Enterobacteriaceae from livestock and poultry as well as their possible role as reservoir of antibiotic resistance genes (ARGs).Methods: Polymerase chain reaction (PCR) and DNA sequencing analyses were used for the characterization of Tn7-like, associated integrons and ARGs. The antimicrobial resistance profile of the isolates was examined by using disc diffusion test.Results: Three hundred and seventy-eight Tn7-like-positive strains of Enterobacteriaceae were isolated, and included E. coli (128), Proteus(150), K. pneumonia(17), Salmonella(13), M. morganii (21) and A. baumannii(1), wherein high resistance was observed for Trimethoprim/Sulfamethoxazole and Streptomycin, and fifty percent of the strains were multidrug-resistant. Integrons class 2 were detected in all of the isolates and there are high frequency mutation sites especially in 535, a stop mutation. Variable region of class 2 integrons carried same gene cassettes, namely aadA1-sat2-dfrA1. From the 378 isolated strains, we found a new type of Tn7-like on a plasmid, named Tn6765.Conclusions: These findings proved that the Tn7-like can contribute to the horizontal transmission of antibiotic resistant genes in livestock and poultry. As potential vessels for antibiotic resistance genes (ARGs), Tn7-like could not be ignored due to their efficient transfer ability in environments.

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Characterization of Two New Shiga Toxin-Producing Escherichia coli O103-Infecting Phages Isolated from an Organic Farm

Microorganisms ◽

10.3390/microorganisms9071527 ◽

2021 ◽

Vol 9 (7) ◽

pp. 1527

Author(s):

Yujie Zhang ◽

Yen-Te Liao ◽

Alexandra Salvador ◽

Valerie M. Lavenburg ◽

Vivian C.H. Wu

Keyword(s):

Escherichia Coli ◽

Host Range ◽

Shiga Toxin ◽

The United States ◽

Antimicrobial Activities ◽

Broad Host Range ◽

Valuable Insight ◽

Antibiotic Resistant ◽

Organic Farm ◽

Double Stranded Dna

Shiga toxin-producing Escherichia coli (STEC) O103 strains have been recently attributed to various foodborne outbreaks in the United States. Due to the emergence of antibiotic-resistant strains, lytic phages are considered as alternative biocontrol agents. This study was to biologically and genomically characterize two STEC O103-infecting bacteriophages, vB_EcoP-Ro103C3lw (or Ro103C3lw) and vB_EcoM-Pr103Blw (or Pr103Blw), isolated from an organic farm. Based on genomic and morphological analyses, phages Ro103C3lw and Pr103Blw belonged to Autographiviridae and Myoviridae families, respectively. Ro103C3lw contained a 39,389-bp double-stranded DNA and encoded a unique tail fiber with depolymerase activity, resulting in huge plaques. Pr103Blw had an 88,421-bp double-stranded DNA with 26 predicted tRNAs associated with the enhancement of the phage fitness. Within each phage genome, no virulence, antibiotic-resistant, and lysogenic genes were detected. Additionally, Ro103C3lw had a short latent period (2 min) and a narrow host range, infecting only STEC O103 strains. By contrast, Pr103Blw had a large burst size (152 PFU/CFU) and a broad host range against STEC O103, O26, O111, O157:H7, and Salmonella Javiana strains. Furthermore, both phages showed strong antimicrobial activities against STEC O103:H2 strains. The findings provide valuable insight into these two phages’ genomic features with the potential antimicrobial activities against STEC O103.

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Prevalence of Multidrug-Resistant Bacteria from U.S.-Grown and Imported Fresh Produce Retailed in Chain Supermarkets and Ethnic Stores of Davidson County, Tennessee

Journal of Food Protection ◽

10.4315/0362-028x.jfp-16-178 ◽

2017 ◽

Vol 80 (3) ◽

pp. 506-514 ◽

Cited By ~ 11

Author(s):

Siqin Liu ◽

Agnes Kilonzo-Nthenge

Keyword(s):

Pathogenic Bacteria ◽

Fresh Produce ◽

Agricultural Practices ◽

Multidrug Resistant ◽

Leafy Vegetables ◽

Resistant Bacteria ◽

Antibiotic Resistant Bacteria ◽

Antibiotic Resistant ◽

E Coli ◽

Public Health Hazards

ABSTRACTThe aim of this study was to determine whether U.S.-grown and imported fresh produce retailed in ethnic stores and chain supermarkets was a reservoir of antibiotic-resistant bacteria. A total of 360 (129 imported and 231 U.S.-grown) samples of fresh produce were purchased from retail stores and analyzed for Enterobacteriaceae, including three pathogenic bacteria (Escherichia coli O157:H7, Shigella, and Salmonella), using standard methods. Presumptive pathogenic isolates were confirmed using PCR. The mean Enterobacteriaceae counts for imported produce were 6.87 ± 0.15 log CFU/g and 7.16 ± 0.11 log CFU/g in ethnic stores and chain supermarkets, respectively. For U.S.-grown produce, the contamination levels were at 8.35 ± 0.17 log CFU/g and 7.52 ± 0.13 log CFU/g in ethnic stores and chain supermarkets, respectively. Salmonella (0 and 0.3%), Shigella (1.7 and 0.6%), E. coli (3.1 and 1.4%), Enterobacter (9.4 and 8.6%), Klebsiella (6.7 and 0.6%), and Serratia (5.8 and 1.4%) were detected in produce from ethnic stores and chain supermarkets, respectively. None of the samples were positive for E. coli O157:H7. Regarding distribution by produce type, leafy vegetables had a significantly (P < 0.05) higher prevalence of Enterobacteriaceae (19.2%) than the other types, followed by root vegetables (6.4%), tomatoes (5.6%), and fruits (3.9%). Antibiotic-resistant Salmonella, Shigella, E. coli, Enterobacter, Klebsiella, and Erwinia bacteria were also isolated from fresh produce. The frequencies of vancomycin resistance (98.1 and 100%) were significantly higher (P < 0.05) than the frequencies of ampicillin resistance (42.3 and 72.9%) for imported and U.S.-grown produce, respectively. Despite the increased attention to the role of imported produce as a source of antimicrobial resistance, this study indicates that U.S.-grown produce is also contaminated with antibiotic-resistant bacteria. Good agricultural practices on the farms and washing of fresh produce before consumption are greatly recommended to avoid possible public health hazards.

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Presence of Multidrug-Resistant Shiga Toxin–Producing Escherichia coli, Enteropathogenic Escherichia coli, and Enterotoxigenic Escherichia coli on Fresh Cheeses from Local Retail Markets in Mexico

Journal of Food Protection ◽

10.4315/0362-028x.jfp-18-166 ◽

2018 ◽

Vol 81 (11) ◽

pp. 1748-1754 ◽

Cited By ~ 1

Author(s):

MARÍA C. de la ROSA-HERNÁNDEZ ◽

ARTURO CADENA-RAMÍREZ ◽

ALEJANDRO TÉLLEZ-JURADO ◽

CARLOS A. GÓMEZ-ALDAPA ◽

ESMERALDA RANGEL-VARGAS ◽

...

Keyword(s):

Escherichia Coli ◽

Shiga Toxin ◽

Pathogenic Bacteria ◽

Multidrug Resistant ◽

Enterotoxigenic Escherichia Coli ◽

Health Concern ◽

Public Markets ◽

E Coli ◽

Unpasteurized Milk ◽

Fresh Cheese

ABSTRACT Cheesemaking is one of the most important industries in Mexico. Among all the Mexican cheeses, fresh cheeses are the most popular and most consumed cheese in Mexico and Latin America. However, in Mexico fresh cheese is frequently made with unpasteurized milk and sold in public markets. This may increase the risk for contamination of dairy products with pathogenic bacteria. The presence of multidrug-resistant pathogenic bacteria in food is an important public health concern. Diarrheagenic Escherichia coli pathotypes (DEPs) are foodborne bacteria. This study investigated the presence of indicator bacteria and multidrug-resistant DEPs in fresh cheeses. A total of 120 fresh cheese samples were collected from public markets in the city of Pachuca, Mexico. The samples were analyzed for presence of fecal coliforms (FC), E. coli, and antibiotic resistant DEPs. FC and E. coli were analyzed using the most-probable-number technique. DEPs were identified using two multiplex PCR methods. Susceptibility to 16 antibiotics was tested for the isolated DEPs strains by the standard assay. The frequency of FC, E. coli, and DEPs in the cheese samples was 50, 40, and 19%, respectively. The identified DEPs included Shiga toxin–producing E. coli (STEC; 8%), enteropathogenic E. coli (EPEC; 6%), and enterotoxigenic E. coli (ETEC; 5%). All isolated strains exhibited resistance to at least five antibiotics. One, one, two, and three STEC strains were resistant to 14, 12, 11, and 10 antibiotics, respectively. One strain of EPEC was resistant to 11 antibiotics, three EPEC strains to 9, and one strain to 7. One, one, and two strains of ETEC were resistant to 10, 8, and 7 antibiotics, respectively. The results of the present study indicate that fresh cheeses made with unpasteurized milk could be a risk for consumers, both for native people and visitors to Mexico.

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Longitudinal Shedding Patterns and Characterization of Antibiotic Resistant E. coli in Pastured Goats using a Cohort Study

Antibiotics ◽

10.3390/antibiotics8030136 ◽

2019 ◽

Vol 8 (3) ◽

pp. 136 ◽

Cited By ~ 1

Author(s):

Ndegwa ◽

Almehmadi ◽

Kim ◽

Kaseloo ◽

Ako

Keyword(s):

Pathogenic Bacteria ◽

Age Groups ◽

Resistant Bacteria ◽

Human Pathogens ◽

Phylogenetic Groups ◽

Antibiotic Resistant ◽

E Coli ◽

Age Cohort ◽

One Year

There is a scarcity of information on antibiotic resistance in goats. To understand shedding of resistant Escherichia coli in pastured goats, we collected fecal samples from a mixed age cohort over a one-year period. No antibiotic had been used on the study animals one year prior to and during the study period. Resistant isolates were detected in all age groups and prevalence in goat kids was significantly higher than adults; 43–48% vs 8–25% respectively. The proportion of resistant isolates was higher when animals were congregated near handling facility than on pasture. Most isolates were resistant to tetracycline (51%) and streptomycin (30%), but also to antibiotics that had never been used on the farm; ampicillin (19%). TetB, bla-TEM, (aadA and strpA/strpB) genes were detected in 70%, 43%, (44% and 24%) of tetracycline, ampicillin, and streptomycin resistant isolates respectively. Resistant isolates also harbored virulent genes and some belonged to D and B2 phylogenetic groups. Thus, pastured goats, despite minimal exposure to antibiotics, are reservoirs of resistant E. coli that may contaminate the environment and food chain and spread resistant genes to pathogenic bacteria and some that are potential animal and human pathogens. Environmental sources may play a role in acquisition of resistant bacteria in pastured goats.

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